The Mahurat for the Official Inauguration of our Assisted Reproduction Center
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Rotunda Walk-thru.mpg - Video
Category Archives: Human Reproduction
"Sex Cells" author on Reproduction Industry – Video
Yale sociologist Rene Almeling, author of "Sex Cells: The Medical Market for Eggs and Sperm" talks here about four important aspects of the unregulated industry in assisted human reproduction: "What defines 'motherhood'?" "Gender differences in claiming parenthood." "Legal ramifications of surrogacy and other assisted reproduction practices." "The lack of oversight of the gamete industry."
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"Sex Cells" author on Reproduction Industry - Video
Rotunda-The Center for Human Reproduction Virtual Walk-thru Video Tour.mpg
The Mahurat for the Official Inauguration of our Assisted Reproduction Center
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Rotunda-The Center for Human Reproduction Virtual Walk-thru Video Tour.mpg
Jack pod cast 4 – Video
Jack's update on human reproduction, age 7.
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Jack pod cast 4 - Video
Human Reproduction – Video
9014, 1940s, DOCTOR EXPLAINS HUMAN REPRODUCTION AND BIRTH TO WOMAN PATIENT, GRAPHIC ILLUSTRATIONS OF REPRODUCTIVE SYSTEM, AND HUMAN BIRTH., http://www.myfootage.com
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Human Reproduction - Video
human reproduction
human reproduction is always sought after by all men, although the smell is not pleasant ... haaaa
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human reproduction
Improvements in human sperm quality by long-term in vitro co-culture with isolated porcine Sertoli cells
BACKGROUND
Spermatogenesis is a complex process where spermatogonial germ cells become spermatozoa with the indispensable support of Sertoli cells (SCs), which provide ‘ad hoc’ structural and nutritional support. Unfortunately, for most sperm dysfunctions, no therapies are yet available except assisted reproductive technologies (ART) that are based on the use of different culture media to preserve sperm in vitro. However, sperm culture is only possible for short periods of time, since long-term culture would invariably and irreversibly damage the cells with negative impact on their fertilization potential.
METHODS
Fresh sperm cells (5 ml of 20 x 106/ml) were co-cultured with SCs layers, derived from prepubertal pig testes or incubated in cell free SC medium or BWW (Biggers, Whitten and Whittingham) medium for 2, 4 or 7 days. Sperm viability, motility, mitochondrial status, DNA fragmentation, chromatin integrity, intracellular calcium and acrosome status were assessed after every co-culture or incubation time, but capacitation and induction of acrosome reaction (AR) with progesterone was only evaluated after 7 days.
RESULTS
SCs layers derived from prepubertal pig testes (co-culture of sperm and SC feeder, CCSCF) were able to preserve normal sperm viability, motility and normal mitochondrial function, after 7 days of culture; CCSCF did not induce AR or hyperactivation of spermatozoa, keeping the sperm in a quiescent state for 7 days of culture. Nevertheless, the sperm were readily able to initiate AR after stimulation with progesterone.
CONCLUSIONS
CCSCF maintained good sperm viability and motility for 7 days. This approach could improve retention of sperm viability and motility during ART procedures and maintain sperm viability, during transfer between two distant Centres, avoiding the need for cryopreservation.
Spore: human male
PLEASE READ DESCRIPTION BEFORE RATING OR COMMENTING! This video was created using Spore. i made a realistic human, reproductive organs and all.
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Spore: human male
Bilateral histological evaluation of exocrine testicular function in men with obstructive azoospermia: condition of spermatogenesis and andrological implications?
BACKGROUND
The aim of this study was to investigate whether there is a need for diagnostic biopsies in men with obstructive azoospermia (OA).
METHODS
Sixty-three adult men with OA due to vasectomy, bilateral inflammation or bilateral aplasia of the vas deferens were included in the study. We determined testicular volume, sexual hormone levels and testicular histologies of right and left testes (236 biopsies from 118 testes) during diagnostic and therapeutic infertility surgery (microsurgical vasal reconstruction or testicular/epididymal sperm extraction). Spermatogenesis was histologically classified according to the Holstein score from 0 (Sertoli cell-only, complete absence of germ cells) to 10 (100% of tubules with elongated spermatids).
RESULTS
All patients (mean age 34 ± 5 years) had low glucosidase levels (5.4 ± 4.2 mU/ejaculate), normal serum FSH levels (4.6 ± 2.5 mU/ml) and normal testicular volumes (right 21 ± 8 ml; left 19 ± 6 ml). Median histological score for right and left testis was 9. There were eight patients with score differences ≥ 3 between right and left testis (14% of men), showing that even in men with OA, there may be differences in spermatogenic activity between both sides. In all of these patients, normal spermatogenesis was found in the larger testis. Testicular histology (spermatogenesis score) was positively correlated with testicular volume and negatively correlated with FSH levels.
CONCLUSIONS
Patients with OA may not need to be biopsied for diagnostic purposes. Our data support the use of unilateral therapeutic biopsy in men with OA and that the larger testicle should be operated on when there is a significant difference in size.
Sodium influx induced by external calcium chelation decreases human sperm motility
BACKGROUND
Calcium removal from the medium promptly reduces human sperm motility and induces a Na+-dependent depolarization that is accompanied by an increase in intracellular sodium concentration ([Na+]i) and a decrease in intracellular calcium concentration ([Ca2+]i). Sodium loading activates a Na+/K+-ATPase.
METHODS
Membrane potential (Vm) and [Ca2+]i were simultaneously detected in human sperm populations with the fluorescent probes diSC3(5) and fura 2. [Na+]i and was measured independently in a similar fashion using sodium-binding benzofuran isophthalate. Motility was determined in a CASA system, ATP was measured using the luciferin-luciferase assay, and cAMP was measured by radioimmunoassay.
RESULTS
Human sperm motility reduction after calcium removal is related to either Na+-loading or Na+-dependent depolarization, because, under conditions that inhibit the calcium removal-induced Na+-dependent depolarization and [Na+]i increase, sperm motility was unaffected. By clamping sperm Vm with valinomycin, we found that the motility reduction associated with the calcium removal was related to sodium loading, and not to membrane potential depolarization. Mibefradil, a calcium channel blocker, markedly inhibited the Na+-dependent depolarization and sodium loading, and also preserved sperm motility. In the absence of calcium, both ATP and cAMP concentrations were decreased by 40%. However ATP levels were unchanged when calcium removal was performed under conditions that inhibit the calcium removal-induced Na+-dependent depolarization and [Na+]i increase.
CONCLUSIONS
Human sperm motility arrest induced by external calcium removal is mediated principally by sodium loading, which would stimulate the Na+/K+-ATPase and in turn deplete the ATP content.
Mast cell tryptase stimulates production of decorin by human testicular peritubular cells: possible role of decorin in male infertility by interfering with growth factor signaling
BACKGROUND
Myofibroblastic, peritubular cells in the walls of seminiferous tubules produce low levels of the extracellular matrix (ECM) protein decorin (DCN), which has the ability to interfere with growth factor (GF) signaling. In men with impaired spermatogenesis, fibrotic remodeling of these walls and accumulation of tryptase-positive mast cells (MCs) occur.
METHODS
Human testicular biopsies with normal and focally impaired spermatogenesis (mixed atrophy) were subjected to immunohistochemistry and laser micro-dissection followed by RT–PCR. Primary human testicular peritubular cells (HTPCs), which originate from normal and fibrotically altered testes (HTPC-Fs), were studied by qRT–PCR, western blotting, enzyme-linked immunosorbent assay measurements and Ca2+ imaging. Phosphorylation and viability/proliferation assays were performed.
RESULTS
Immunohistochemistry revealed DCN deposits in the walls of tubules with impaired spermatogenesis. Mirroring the situation in vivo, HTPC-Fs secreted more DCN than HTPCs (P< 0.05). In contrast to HTPCs, HTPC-Fs also responded to the main MC product, tryptase, and to a tryptase receptor (PAR-2) agonist by further increased production of DCN (P< 0.05). Several GF receptors (GFRs) are expressed by HTPCs and HTPC-Fs. DCN acutely increased intracellular Ca2+-levels and phosphorylated epidermal GF (EGFR) within minutes. Platelet-derived GF (PDGF) and EGF induced strong mitogenic responses in HTPC/-Fs, actions that were blocked by DCN, suggesting that DCN in the ECM interferes with GF/GFRs signaling of peritubular cells of the human testis.
CONCLUSIONS
The data indicate that the increase in testicular DCN found in male infertility is a consequence of actions of MC-derived tryptase. We propose that the increases in DCN may consequently imbalance the paracrine signaling pathways in human testis.
Recurrent pre-clinical pregnancy loss is unlikely to be a ’cause’ of unexplained infertility
BACKGROUND
A proportion of women with ‘unexplained’ infertility may present with subfertility because their pregnancies fail before they are clinically recognized. In order to test whether pre-clinical early pregnancy losses (EPL) occur more frequently in women with unexplained infertility, serial urinary hCG concentrations were measured to compare EPL per cycle rates following spontaneous conception in patients with unexplained infertility versus healthy volunteers.
METHODS
Sixty patients under 39 years of age with unexplained infertility and 60 healthy controls, who were trying to conceive spontaneously, participated in this study. All participants were asked to collect daily urine samples from cycle day 14 until menstruation for three consecutive cycles or until a positive pregnancy test was obtained.
Urinary hCG and creatinine levels were measured by immunoassay. Implantation was detected when urinary hCG levels rose above reference levels constructed from samples obtained from 12 women not attempting to conceive. EPL rates were determined by a linear mixed model using logarithmically transformed hCG/creatinine data.
RESULTS
In the 133 cycles of 60 women with unexplained infertility, just one implantation was detected, which became an ongoing pregnancy. In contrast, in 103 such cycles in 46 control patients, 30 implantations were detected (24 clinical pregnancies, 6 cases of EPL). The odds ratio for EPL/cycle in the unexplained versus control group was 0 (95% confidence interval: 0–0.795, P= 0.026).
CONCLUSIONS
Our data do not support the hypothesis that recurrent EPL may present as unexplained infertility. Post-implantation failure is therefore unlikely to contribute significantly to the presentation of subfertility.
Influence of BMI on risk of miscarriage after single blastocyst transfer
BACKGROUND
Debate exists regarding the effect of raised BMI on the outcome of pregnancies after assisted reproduction technology. We assessed the effect of BMI on the risk of miscarriage in women conceiving following single blastocyst transfer (SBT) after controlling for confounding factors.
METHODS
Fresh and cryo-thawed cycles of SBT that resulted in a pregnancy between January 2006 and March 2010 were included. Patients with BMI < 18.5 kg/m2 or older than 40 years were excluded. Patients were grouped according to their BMI at the start of treatment cycle. The main outcome measure was the miscarriage rate before 23 weeks gestation. Confounding variables examined included female age, duration and cause of infertility, previous miscarriage, smoking status and quality of blastocyst replaced.
RESULTS
A total of 413 women conceived following SBT in fresh (n= 325) or cryo-thawed (n= 88) IVF cycles, of whom 244 had a normal BMI (18.5–24.9) and 169 had a raised BMI of ≥25. Overall, 27% (113/413) of women miscarried before 23 weeks gestation. Women with a BMI of ≥25 had more than double the risk of miscarriage compared with women who had normal BMI [38 versus 20%, odds ratio (OR): 2.4, 95% confidence interval (CI) 1.6–3.8, P < 0.001, respectively]. After adjusting for confounding variables, having a BMI of ≥25 significantly increased the risk of clinical miscarriage before 23 weeks gestation in both fresh (adjusted OR = 2.7, 95% CI 1.5–4.9, P= 0.001) and cryo-thawed IVF cycles (OR = 6.8, 95% CI 1.5–31.1, P= 0.012).
CONCLUSIONS
Raised BMI is independently associated with higher miscarriage rate after IVF treatment.
Contribution of referent pathologists to the quality of trophoblastic diseases diagnosis
OBJECTIVE
To evaluate the contribution of referent pathologists (RPs) to the quality of diagnosis of trophoblastic diseases and to study the level of diagnostic agreement between the initial pathologists and the RPs.
METHODS
This observational retrospective study was carried between 1 November 1999 and 11 January 2011 using the database of the French Trophoblastic Disease Reference Centre in Lyon. All files for hydatiform moles (HMs), trophoblastic tumours and non-molar pregnancies for which there was an initial suspicion of trophoblastic disease were included, whenever there was rereading of the slides by an RP. A total of 1851 HMs and 150 gestational trophoblastic tumours were analysed.
RESULTS
When the initial pathologist diagnosed a complete mole, the RP confirmed the diagnosis in 96% of cases. When the initial pathologist diagnosed a partial mole, the RP confirmed the diagnosis in only 64% of cases. For trophoblastic tumours, when the initial pathologist diagnosed a choriocarcinoma, the RP confirmed the diagnosis in 86% of cases. When the initial anatomopathology suggested an invasive mole, the diagnosis was confirmed in 96% of cases. Finally, when the initial diagnosis was a placental site trophoblastic tumour or an epithelioid trophoblastic tumour, the RP confirmed the diagnosis in 60 and 100% of cases, respectively.
CONCLUSION
A systematic policy of rereading of slides for all suspicious moles improves the quality of management of trophoblastic diseases at a national level.
3D ultrasound to assess the position of tubal sterilization microinserts
BACKGROUND
The aim of this study was to assess the diagnostic accuracy of three-dimensional ultrasound (3D-US) for determining the position of Essure microinserts and the success of sterilization by the Essure method.
METHODS
This retrospective observational study examined the case records of 311 women who underwent hysteroscopic sterilization from October 2002 through October 2008. Imaging with 3D-US or pelvic X-radiography or both was performed 3 months after the procedure to verify device position. Hysterosalpingography (HSG) was performed when a bilateral procedure was not completed because of a history of salpingectomy or blocked tube, when doubt persisted after 3D-US or pelvic radiography, or for comparative purposes in a prospective study. The positions seen on 3D-US were classified in four categories according to a specific scale we devised.
RESULTS
The insertion procedure was completed in 94.2% patients. Only 90.5% underwent imaging verification of the device 3 months afterwards. In all, 227 3D-US, 175 pelvic radiography and 64 HSG imaging procedures were performed. Visualization of the device was possible in 99.6% of the 3D-US images. According to our classification, 3D-US was appropriate for assessing device position for 195 (85.9%) patients. The need for HSG confirmation was significantly lower with 3D-US than radiographic imaging (14.1 versus 26.8%, P = 0.001). 3D-US examinations, compared with the results of HSG as the reference test, had a sensitivity of 100% and a specificity of 76.6%. Neither pregnancy nor early expulsion occurred when 3D-US found that the devices were correctly placed.
CONCLUSIONS
3D-US is a simple technique for assessing the position of Essure® microinserts, even after concomitant endometrial surgery. The 3D-US classification presented here appears to make it possible to use HSG for back-up confirmation only when the microinsert is found in a very distal position on 3D-US and thus to protect the majority of women from the negative effects of pelvic radiography and HSG.
Analysis of global gene expression following mouse blastocyst cryopreservation
BACKGROUND
The aim of this study was to examine the effect of the cryopreservation procedure (slow freezing or vitrification) and cryoprotectants (1,2-propanediol or dimethylsulphoxide) on mouse blastocyst gene expression.
METHODS
Cultured mouse blastocysts were cryopreserved with different protocols. Following thawing/warming, total RNA from re-expanded blastocysts was isolated, amplified and then analyzed using mouse whole-genome microarrays.
RESULTS
Compared with non-cryopresevered control blastocysts, gene expression was only significantly altered by slow freezing. Slow freezing affected the expression of 115 genes (P < 0.05). Of these, 100 genes exhibited down-regulation and 15 genes were up-regulated. Gene ontology revealed that the majority of these genes are involved in protein metabolism, transcription, cell organization, signal transduction, intracellular transport, macromolecule biosynthesis and development. Neither of the vitrification treatment groups showed statistically different gene expression from the non-cryopreserved control embryos. Hierarchical cluster analysis, did however, reveal that vitrification using 1,2-propanediol could result in a gene expression profile closest to that of non-cryopreserved blastocysts.
CONCLUSIONS
Investigating the effects of cryopreservation on cellular biology, such as gene expression, is fundamental to improving techniques and protocols. This study demonstrates that of the cryopreservation regimens employed, slow freezing induced the most changes in gene expression compared with controls.
The use of morphokinetics as a predictor of embryo implantation
BACKGROUND
Time-lapse observation presents an opportunity for optimizing embryo selection based on morphological grading as well as providing novel kinetic parameters, which may further improve accurate selection of viable embryos. The objective of this retrospective study was to identify the morphokinetic parameters specific to embryos that were capable of implanting. In order to compare a large number of embryos, with minimal variation in culture conditions, we have used an automatic embryo monitoring system.
METHODS
Using a tri-gas IVF incubator with a built-in camera designed to automatically acquire images at defined time points, we have simultaneously monitored up to 72 individual embryos without removing the embryos from the controlled environment. Images were acquired every 15 min in five different focal planes for at least 64 h for each embryo. We have monitored the development of transferred embryos from 285 couples undergoing their first ICSI cycle. The total number of transferred embryos was 522, of which 247 either failed to implant or fully implanted, with full implantation meaning that all transferred embryos in a treatment implanted.
RESULTS
A detailed retrospective analysis of cleavage times, blastomere size and multinucleation was made for the 247 transferred embryos with either failed or full implantation. We found that several parameters were significantly correlated with subsequent implantation (e.g. time of first and subsequent cleavages as well as the time between cleavages). The most predictive parameters were: (i) time of division to 5 cells, t5 (48.8–56.6 h after ICSI); (ii) time between division to 3 cells and subsequent division to 4 cells, s2 (≤0.76 h) and (iii) duration of cell cycle two, i.e. time between division to 2 cells and division to 3 cells, cc2 (≤11.9 h). We also observed aberrant behavior such as multinucleation at the 4 cell stage, uneven blastomere size at the 2 cell stage and abrupt cell division to three or more cells, which appeared to largely preclude implantation.
CONCLUSIONS
The image acquisition and time-lapse analysis system makes it possible to determine exact timing of embryo cleavages in a clinical setting. We propose a multivariable model based on our findings to classify embryos according to their probability of implantation. The efficacy of this classification will be evaluated in a prospective randomized study that ultimately will determine if implantation rates can be improved by time-lapse analysis.
Effective treatment of heavy and/or prolonged menstrual bleeding with an oral contraceptive containing estradiol valerate and dienogest: a randomized, double-blind Phase III trial
BACKGROUND
This double-blind trial investigated the efficacy and safety of estradiol valerate/dienogest (E2V/DNG) for the treatment of heavy menstrual bleeding without recognizable organic pathology.
METHODS
Otherwise healthy women with idiopathic heavy, prolonged or frequent menstrual bleeding, confirmed during a 90-day run-in phase, were randomized (2:1) according to a permuted-block, computer-generated schedule to E2V/DNG or placebo for 196 days at 34 centres in Europe and Australia. The primary efficacy end-point was the proportion of women with a ‘complete’ response (i.e. a return to ‘menstrual normality’) during a 90-day efficacy phase. Secondary end-points included changes in measured menstrual blood loss (MBL) and iron metabolism parameters.
RESULTS
The intention-to-treat population comprised 231 women. The E2V/DNG response rate was much higher than with placebo (P < 0.0001). The mean reduction in MBL volume in E2V/DNG recipients was 69.4% (median 79.2%) versus 5.8% (median 7.4%) in placebo recipients. The between-treatment difference in MBL volume was 373 ml in favour of E2V/DNG (95% confidence interval 490, 255 ml; P < 0.0001). Significant improvements in iron metabolism parameters were observed with E2V/DNG but not placebo. Overall, 14 women (9.7%) treated with E2V/DNG and 5 (6.2%) treated with placebo prematurely discontinued treatment because of adverse events, headache being the most prevalent. Serious adverse events occurred in both the E2V/DNG and placebo groups (each n = 2).
CONCLUSIONS
E2V/DNG is an effective treatment in women with heavy and/or prolonged menstrual bleeding without organic pathology. Further study of E2V/DNG compared with an active comparator is warranted.
ClinicalTrials.gov identifier: NCT00307801.
One- and two-day dosing intervals between mifepristone and misoprostol in second trimester medical termination of pregnancy–a randomized trial
BACKGROUND
The recommended time interval between mifepristone and misoprostol in medical second trimester termination of pregnancy (TOP) has been 36–48 h. However, a more flexible interval would be of value. The aim of this investigation was to compare one- and two-day intervals in second trimester medical TOP. The main outcome measures were induction-to-abortion time and the rate of surgical evacuation.
METHODS
This open randomized trial included 227 women undergoing TOP between gestational weeks 13–24. Mifepristone (200 mg) was followed by misoprostol (400 mcg) after one (17–28 h) or two (41– 45 h) days.
RESULTS
In intention-to-treat analysis, the median induction-to-abortion interval was 1h longer in the one-day group (8.5 versus 7.2 h, P= 0.038), but in per-protocol analysis, the rate of surgical evacuation was higher in the 2-day group [30/115 (25%) versus 40/112 (37%); 95% confidence interval 0.3–24.1, P= 0.044]. A subgroup analysis showed that the median induction-to-abortion interval was 3h longer in the one-day group, amongst women without previous vaginal deliveries (10.1 versus 7.6, P= 0.013) and when gestation exceeded 16 weeks (10.8 versus 7.2, P= 0.024).
CONCLUSIONS
Both one- and two-day dosing intervals seem to be suitable for second trimester medical TOP, but women with no previous deliveries and those whose gestation exceeds 16 weeks may benefit from the longer interval. However, evaluated on the basis of surgical evacuation, the one-day interval could be supported as an option for second trimester medical TOP. Effective use of both one- and two-day dosing intervals is important when optimizing clinical service.
Trial Registration: ISRCTN09944151.
Lymphangiogenesis in deep infiltrating endometriosis
BACKGROUND
In patients diagnosed with deep infiltrating endometriosis (DIE), foci of endometriosis are detected in mesorectal lymph nodes (LNs) after segmental bowel resection and in pelvic sentinel LNs. Lymph vessels (LVs) seem to be the possible routes for the dissemination of endometriotic cells from DIE-lesions to LN. Therefore, we conducted a study to investigate the occurrence and density of LV and lymphangiogenic growth factors in DIE.
METHODS
Included in this study were 38 premenopausal women who underwent surgery due to symptomatic rectovaginal DIE. In order to identify LV, immunohistochemical analysis with anti-Podoplanin (D2-40), LYVE-1 and Prox-1 was performed. Furthermore, the expression of VEGF-C and VEGF-D in endometriotic tissue was investigated.
RESULTS
LV density (LVD) of DIE lesions was significantly higher compared with healthy corresponding tissue. All LV makers could be detected, and the density of LYVE-1- or Prox-1-positive LV was significantly higher than that of D2-40-positive LV. Endometriotic epithelial cells and stromal cells showed a moderate to strong VEGF-C and VEDF-D expression.
CONCLUSIONS
DIE lesions have lymphangiogenic properties, probably leading to endometriosis-like cells in lymphatic vessels and LNs featuring a loco-regional disease.